WO2017173347A1 - Peptides mimétiques par2 et utilisations desdits peptides - Google Patents

Peptides mimétiques par2 et utilisations desdits peptides Download PDF

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WO2017173347A1
WO2017173347A1 PCT/US2017/025511 US2017025511W WO2017173347A1 WO 2017173347 A1 WO2017173347 A1 WO 2017173347A1 US 2017025511 W US2017025511 W US 2017025511W WO 2017173347 A1 WO2017173347 A1 WO 2017173347A1
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par
composition
moiety
gly
ile
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PCT/US2017/025511
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Scott A. BOITANO
Josef Vagner
Theodore J. Price
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Arizona Board Of Regents On Behalf Of The University Of Arizona
Board Of Regents, The University Of Texas System
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Priority to US16/090,525 priority Critical patent/US11351264B2/en
Publication of WO2017173347A1 publication Critical patent/WO2017173347A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
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    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
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    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
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    • A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
    • A—HUMAN NECESSITIES
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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/545—Heterocyclic compounds
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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/081—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/101—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/1013—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1024—Tetrapeptides with the first amino acid being heterocyclic
    • C—CHEMISTRY; METALLURGY
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6427—Chymotrypsins (3.4.21.1; 3.4.21.2); Trypsin (3.4.21.4)
    • C—CHEMISTRY; METALLURGY
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
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    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6445—Kallikreins (3.4.21.34; 3.4.21.35)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention is in the field of medicinal pharmacology.
  • the invention relates to protease activated receptor type 2 (PAR 2 ) modulating compounds (e.g., mimetic peptides), compositions comprising such modulating compounds, and their use as therapeutics for the treatment of conditions involving PAR 2 activity.
  • PAR 2 protease activated receptor type 2
  • Asthma is a growing and potentially debilitating disease in the industrialized world.
  • Migraine pain is a major clinical problem. Almost 15 percent of the global population is affected by migraines during their lifetimes (see, e.g., Vos, T., et al, Lancet, 2012. 380(9859): p. 2163-96), and there are over 36 million migraine sufferers in the US alone. Even with this significant number of patients, treatments for migraine pain remains little more effective than over- the-counter analgesics. Part of the problem is that migraine etiology is complex and not well understood. Unlike common headaches, migraines have a specific presentation in which a prodrome, aura, and postdrome may occur with the migraine pain lasting between 4 and 72 hours.
  • SLIGRL cortical spreading depression
  • NO nitric oxide
  • CGRP calcitonin gene-related peptide
  • SLIGRL is problematic as it also activates MrgprCl 1, a receptor that is expressed in DRG and TG neurons and contributes to sensory neuron sensitization, with overlapping potency and efficacy to SLIGRL action at PAR 2 (see, e.g., Ramachandran, R. and M.D. Hollenberg, Br J Pharmacol, 2008. 153 Suppl 1 : p.
  • Protease-activated receptor type 2 is a G-protein-coupled receptor (GPCR) implicated in disease conditions including allergic asthma (Br J Pharmacol 2009;158: 1017-33), cancer (Scand J Gastroenterol 2008;43:902-9) arthritis (Biol Chem 2008;389:971-82), and chronic pain (Physiol Rev 2004;84:579-621)
  • GPCR G-protein-coupled receptor
  • PAR 2 can be activated in response to various exogenous and endogenous proteases (Br J Pharmacol 2008; 153(suppl l ):S263-282).
  • mice have been indispensable for elucidating the role of this receptor in normal physiology and pathology (Physiol Rev 2004;84:579-621 ), a lack of suitable pharmacological tools have hindered full exploration of the role of this receptor in disease conditions, including chronic pain (Pharmacol Ther 2011;130:248-82).
  • Highly potent, efficacious, and specific agonists have been developed (J Med Chem 2011 ; 54: 1308-13; J Biol Chem 2011 ; 286: 19076-88; J Physiol 2007; 578:715-33) and used them in experiments to explore the role of PAR2 in the development of a chronic pain state.
  • a central hypothesis for experiments conducted during the course of developing embodiments for the present invention was that PAR 2 plays a pivotal role in causing acute pain, promoting chronic pain, and in both promoting and controlling asthma symptoms, and that high affinity ligands of PAR 2 will represent a novel class of analgesics with utility in a number of chronic pain conditions and in the control of asthma.
  • a primary objective of experiments conducted during the course of developing embodiments for the present invention was to develop novel and specific ligands to PAR 2 , to fully elucidate PAR 2 contribution to acute and chronic pain and asthma, and to evaluate PAR 2 ligand efficacy as novel analgesics in preclinical pain and asthma models.
  • the present invention relates to modulating compounds that function as activators and inhibitors of PAR 2 proteins.
  • the invention further relates to methods of treating, ameliorating, or preventing disorders in a patient, such as those that are responsive to either PAR 2 activation or PAR 2 inhibition, comprising administering to a subject (e.g., a human patient) a composition comprising one or more of the PAR 2 modulating compounds of the invention and, potentially, additional agent(s).
  • disorders include those characterized by aberrant PAR 2 activity (e.g., inflammatory disorders such as asthma and chronic pain).
  • the present invention is not limited to particular types or kinds of modulating compounds that function as activators and inhibitors of PAR 2 proteins.
  • the modulating compounds include small molecule compounds and mimetic peptides.
  • the modulating compounds which function as activators and inhibitors of PAR 2 proteins are PAR 2 mimetic peptides.
  • the present invention provides compositions comprising a PAR 2 mimetic peptide.
  • the PAR 2 mimetic peptide is encompassed within Formula I:
  • Such PAR-2 mimetic peptides are not limited to a particular heterocycle moiety.
  • the heterocycle moiety comprises at least one atom selected from Nitrogen, Oxygen and Sulfur.
  • the heterocycle moiety is selected from the group consisting of a thiazole moiety, a pyridine moiety, an azabicycloalkane moiety, an aminothiazoyl moiety, and an aminonicotinyl moiety.
  • Such PAR-2 mimetic peptides are not limited to a particular position for the heterocycle moiety.
  • the heterocycle moiety is positioned at the N-terminus of the PAR 2 mimetic peptide.
  • Such PAR 2 mimetic peptides are not limited to a particular peptide sequence.
  • the peptide sequence comprises two or more contiguous amino acid residues.
  • the two or more contiguous amino acid residues render the resulting PAR 2 mimetic peptide as a PAR 2 activator and/or a PAR 2 inhibitor.
  • the amino acid sequence selected from the group consisting of Ile-Gly, Ile-Gly-Arg, Leu-Ile-Gly, Leu-Ile-Gly, Leu-Ile-Gly-Arg, Ser-Leu-Ile-Gly, Ser-Leu-Ile-Gly-Arg, Thr-Ile-Gly, Thr-Ile-Gly-Arg, Ser-Lys- Gly-Arg-Ser, Ser-Lys-Gly-Arg, His-Ile-Gly-Arg, Val-Ile-Gly-Arg, any of the peptide sequences described in Example 1, and any of the peptide sequences described in Tables 1, 2, 3, 4 and 5.
  • the linker moiety comprises a chemical moiety configured to bridge the peptide sequence and cell membrane anchoring moiety.
  • the linker moiety comprises a chemical moiety selected from the group consisting of a substituted aliphatic chain, an unsubstituted aliphatic chain, substituted aromatic moiety, an unsubstituted aromatic moiety, a linear polymer, one or more polyethylene glycol (PEG) moieties, one or more 3,19-dioxo- 2,8,1 l,14,21-pentaoxa-4,18-diazatricosan-23-oic acid residue derivative moieties, and/or any combination thereof.
  • PEG polyethylene glycol
  • the linker moiety comprises a polyethylene glycol (PEG) moiety. In some embodiments, the linker moiety comprises multimers of 3,19-dioxo- 2,8,1 l,14,21-pentaoxa-4,18-diazatricosan-23-oic acid. In some embodiments, the linker moiety comprises a polyethylene glycol (PEG) moiety. In some embodiments, the linker moiety is a linear polymer comprising monomelic subunits. In some embodiments, the linear polymer comprises saccharide moieties, peptide moieties, lactone moieties, acrylate moieties, and/or synthetic polymer moieties.
  • the linear polymer comprises collagen-like polypeptides and/or synthetic surrogates of spider silk.
  • Such PAR-2 mimetic peptides are not limited to a particular cell membrane anchoring moiety.
  • the cell membrane anchoring moiety comprises a hydrophobic chemical moiety or a synthetic structure that forms a non-covalent binding interaction with a cell membrane.
  • the cell membrane anchoring moiety is positioned at the C-terminus of the PAR2 mimetic peptide.
  • the cell membrane anchoring moiety comprises a lipid moiety.
  • the cell membrane anchoring moiety comprises a saturated or unsaturated hydrocarbon moiety.
  • the cell membrane anchoring moiety is hexadecyl.
  • the cell membrane anchoring moiety is a saturated C 12 - C 2 0 alkyl residue.
  • the cell membrane anchoring moiety is a cell membrane homing structure.
  • the cell membrane anchoring moiety is a cell-penetrating moiety.
  • the cell membrane anchoring moiety is a transmembrane domain.
  • the PAR 2 mimetic peptide is configured to modulate PAR 2 biological activity.
  • the PAR 2 mimetic peptide is configured to activate PAR 2 biological activity.
  • the mimetic peptide is: [2- aminothiazoyl]-[LIGR]- Pi?G 3 ]-[hexadecyl]. In some such embodiments, the mimetic peptide is:
  • the mimetic peptide is: [2-aminothiazoyl]-[VIGR]- P£ , G 3 ]-[hexadecyl]. In some such embodiments, the mimetic peptide is: [2-aminothiazoyl]-[(homoserine)IGR]- P£ , G 3 ]- [hexadecyl].
  • the PAR 2 mimetic peptide is configured to antagonize PAR 2 biological activity.
  • the mimetic peptide is configured to antagonize PAR 2 activity resulting from interaction between trypsin and PAR 2 .
  • the mimetic peptide is: [2-aminothiazoyl]-[Thr-Ile-Gly-Arg]- P£'G3]-[hexadecyl].
  • the mimetic peptide is configured to antagonize PAR 2 activity resulting from interaction between kallikrein 5 and PAR2.
  • the mimetic peptide is: [2-aminothiazoyl]-[Ser-Lys-Gly-Arg-Ser]- P£ , G 3 ]-[hexadecyl].
  • the mimetic peptide is: [2-aminothiazoyl]-[Ser-Lys-Gly-Arg]- P£ , G 3 ]-[hexadecyl].
  • the mimetic peptide is: [2-aminothiazol-4yl]-[SKGRS]-/PEG3]-
  • the mimetic peptide is: [2-aminothiazol-4yl]-[SKGR]-/PEG3]-
  • the mimetic peptide is: [2-aminothiazol-4yl]-[LIGR]-/PEG3] -
  • the mimetic peptide is: [2-aminothiazol-4yl]-[TIGR]-/PEG3] - [Hdc .
  • the mimetic peptide is shown in Tables 1, 2, 3, 4, and/or 5. In some embodiments, the mimetic peptide is 2-at-LIGRL-P£ , G 3 -Hdc (
  • the modulating compounds which function as activators and inhibitors of PAR 2 proteins are small molecules.
  • the present invention rovides small molecule compounds encompassed within Formula II:
  • Formula I is not limited to a particular chemical moiety for R 1; R 2 , R3, and R4.
  • the particular chemical moiety for R 1; R 2 , R3, and R4 independently include any chemical moiety that permits the resulting compound to function as an inhibitor of PAR 2 protein activity.
  • the particular chemical moiety for R 1; R 2 , R 3 , and R4 independently include any chemical moiety that permits the resulting compound to function as an activator of PAR 2 protein activity.
  • Such compounds are not limited to a particular chemical moiety for Ri.
  • R 2 is an amino acid selected from a Leu, He, Val, Cha, Arg, Orn, Lys, Dap, Thr, Ser, and Tyr.
  • R4 is selected from 2-furoyl acetyl ( O ), 3-
  • the modulating compound is the PAR2 antagonist C391 (
  • the C391 is lipidated.
  • the present invention provides methods for modulating the activity of PAR 2 in a subject (e.g., human subject, non-human subject), comprising administering to the subject a PAR 2 modulating compound as described herein (e.g., a mimetic peptide, a small molecule) of the present invention.
  • a PAR 2 modulating compound as described herein (e.g., a mimetic peptide, a small molecule) of the present invention.
  • the subject is experiencing aberrant
  • the subject is at risk for experiencing aberrant PAR 2 activity.
  • the subject has or is at risk for developing an inflammatory condition (e.g., asthma) involving aberrant PAR 2 activity.
  • the subject has or is at risk for developing chronic pain involving aberrant PAR 2 activity.
  • the inflammatory condition is one or more conditions selected from the group consisting of asthma, chronic pain, cancer, and a vascular disorder.
  • the methods further comprise administering to the subject one or more additional agents (e.g., anti-inflammatory agents, anti-cancer agents, pain-relieving agents).
  • the additional agent is an anti-inflammatory agent.
  • the anti-inflammatory agent is a non-steroidal anti -inflammatory drug.
  • antiinflammatory agent is albuterol.
  • the pharmaceutical composition comprises a PAR 2 mimetic peptide of the present invention and a pharmaceutically acceptable carrier.
  • kits comprising a pharmaceutical composition comprising a PAR 2 mimetic peptide of the
  • FIG. 1 shows the primary PAR 2 signalling pathways.
  • FIG. 2 shows a schematic for PAR 2 signalling, and the measuring of PAR 2 signalling.
  • FIG. 3 shows PAR 2 tethered ligand probe development - trypsin site.
  • FIG. 4 shows PAR 2 tethered ligand probe development - kallikrein site.
  • FIG. 5 shows in vitro physiological PAR agonist screening using xCELLigence.
  • FIG. 6 shows 2-at-TIGR-Pi?G3-Hcfc signalling assays. DETAILED DESCRIPTION OF THE INVENTION
  • the protease-activated receptor-2 (PAR 2 ) is one of the four members of the family of GPCRs that are activated after proteolytic cleavage of their extracellular, amino terminus (Adams et al., Pharmacol. Ther. 130, 248-282; Ramachandran, R, et al, (2012) Nat. Rev. Drug Discov. 11, 69-86).
  • the resulting 'tethered-peptide' sequence (ending with SLIGRL in the rodent receptor and SLIGKV in the human receptor) exposed after proteolytic cleavage activates PAR 2 .
  • PAR-2 plays an important role in a variety of diseases linked to proteinase release from endogenous sources or exposure to exogenous proteinases (Ramachandran R, et al., (2012) Nat Rev Drug Discov 11 : 69-86; Hollenberg, M.D., et al, 2014. 171(5): p. 1180-94).
  • One consequence of PAR 2 activation in the peripheral nervous system is sensitization of neurons responsible for transmitting noxious information to the CNS.
  • the present invention provides PAR 2 mimetic peptides that utilize this ligand chemistry combined with alternative PAR 2 cleavage sites. Indeed, experiments conducted during the course of developing embodiments for the present invention identified highly potent PAR 2 peptides and mimetic activators and agonists.
  • the present invention relates to modulating compounds which function as activators and inhibitors of PAR 2 proteins.
  • the invention further relates to methods of treating, ameliorating, or preventing disorders in a patient, such as those that are responsive to either PAR 2 activation or PAR 2 inhibition, comprising administering to a subject (e.g., a human patient) a composition comprising one or more of the PAR 2 mimetic peptides off the invention and, potentially, additional agent(s).
  • disorders include those characterized by aberrant PAR 2 activity (e.g., inflammatory disorders).
  • modulating compounds include mimetic peptides which function as activators and inhibitors of PAR 2 proteins.
  • the present invention provides PAR 2 mimetic peptides having Formula I:
  • Formula I is not limited to particular chemical moieties for the heterocyle moiety, the peptide sequence, the linker moiety, and/or the cell membrane anchoring moiety.
  • the heterocycle moiety is any aromatic heterocycle moiety that comprises at least one atom selected from Nitrogen, Oxygen and Sulfur.
  • heterocyle moieties include, but are not limited to, a thiazole moiety, a pyridine moiety, an azabicycloalkane moiety, an aminothiazoyl moiety, and/or an aminonicotinyl moiety.
  • the heterocyle moiety is not limited to a particular positioning within the PAR 2 mimetic peptide.
  • the aromatic heterocycle moiety is positioned at the N-terminus of the PAR 2 mimetic peptide.
  • the peptide sequence is any peptide sequence that comprises two or more contiguous amino acid residues. In some embodiments, the peptide sequence is any combination of two or more contiguous amino acid residues that confers PAR 2 activating properties or PAR 2 antagonizing properties onto the PAR 2 mimetic peptide.
  • Examples of the two or more contiguous amino acid residues include, but are not limited to Ile-Gly, Ile-Gly-Arg, Leu-Ile-Gly, Leu-Ile-Gly, Leu-Ile-Gly-Arg, Ser-Leu-Ile-Gly, Ser-Leu-Ile-Gly-Arg, Thr-Ile-Gly, Thr-Ile-Gly- Arg, Ser-Lys-Gly-Arg-Ser, SKGR, HIGR, VIGR, any of the peptide sequences described in Example 1, and any of the peptide sequences described in Tables 1, 2, 3, 4 and 5.
  • the linker moiety is a chemical moiety configured to bridge the peptide sequence and cell membrane anchoring moiety.
  • linker moieties include, but are not limited to, a substituted aliphatic chain, an unsubstituted aliphatic chain, substituted aromatic chain, an unsubstituted aromatic chain, a linear polymer, one or more polyethylene glycol (PEG) moieties, one or more 3,19-dioxo-2,8,l l,14,21-pentaoxa-4,18-diazatricosan-23-oic acid residue derivative moieties, and/or any combination thereof.
  • the linker moiety comprises a polyethylene glycol (PEG) moiety.
  • the linker moiety comprises multimers of 3,19-dioxo-2,8,l l,14,21-pentaoxa-4,18-diazatricosan-23-oic acid.
  • the linker moiety is a linear polymer that comprises monomeric subunits.
  • the linear polymer comprises saccharide moieties, peptide moieties, lactone moieties, acrylate moieties, and/or synthetic polymer moieties.
  • the linear polymer comprises collagen-like polypeptides and/or synthetic surrogates of spider silk.
  • the cell membrane anchoring moiety is any chemical moiety that comprises a hydrophobic chemical moiety or a synthetic structure that forms a non-covalent binding interaction with a cell membrane.
  • the PAR2 mimetic peptides are not limited to a particular type or kind of a cell membrane anchoring moiety.
  • cell membrane moiety comprises a lipid moiety.
  • the cell membrane anchoring moiety comprises a saturated or unsaturated hydrocarbon moiety.
  • the cell membrane anchoring moiety is hexadecyl.
  • the cell membrane anchoring moiety is a saturated C 12 - C 20 alkyl residue.
  • the cell membrane anchoring moiety is a cell-penetrating moiety.
  • the cell membrane anchoring moiety is a transmembrane domain.
  • the cell membrane anchoring moiety is not limited to a particular positioning within the
  • the cell membrane anchoring moiety is positioned at the C-terminus of the PAR 2 mimetic peptide.
  • the length of the [linker moiety] -[cell membrane anchoring moiety] is approximately 30 - 50 Angstroms.
  • the PAR 2 mimetic peptide is configured to modulate PAR 2 biological activity.
  • the PAR 2 mimetic peptide is configured to activate PAR 2 biological activity.
  • the mimetic peptide is: [2- aminothiazoyl]-[LIGR]- Pi?G3]-[hexadecyl]. In some such embodiments, the mimetic peptide is:
  • the mimetic peptide is: [2-anrinothiazoyl]-[VIGR]- Pi?G3Hhexadecyl]. In some such embodiments, the mimetic peptide is: [2-aminothiazoyl]-[(homoserine)IGR]- P£ , G 3 ]- [hexadecyl].
  • the PAR 2 mimetic peptide is configured to antagonize PAR 2 biological activity.
  • the mimetic peptide is configured to antagonize PAR 2 activity resulting from interaction between trypsin and PAR 2 .
  • the mimetic peptide is: [2-aminothiazoyl]-[Thr-Ile-Gly-Arg]- P£ , G 3 ]-[hexadecyl].
  • the mimetic peptide is configured to antagonize PAR 2 activity resulting from interaction between kallikrein 5 and PAR2.
  • the mimetic peptide is: [2-aminothiazoyl]-[Ser-Lys-Gly-Arg-Ser]- P£ , G 3 ]-[hexadecyl].
  • the mimetic peptide is: [2-aminothiazoyl]-[Ser-Lys-Gly-Arg]- P£'G3]-[hexadecyl].
  • the mimetic peptide is: [2-aminothiazol-4yl]-[SKGRS]-/PEG 3 ]- [Hdc .
  • the mimetic peptide is: [2-aminothiazol-4yl]-[SKGR]-/PEG 3 ]-
  • the mimetic peptide is: [2-aminothiazol-4yl]-[LIGR]-/PEG 3 ]-
  • the mimetic peptide is: [2-aminothiazol-4yl]-[TIGR]-/PEG 3 ]-
  • the mimetic peptide is shown in Tables 1, 2, 3, 4, and/or 5. In some embodiments, the mimetic peptide is 2-at-LIGRL-P£G 3 -Hdc (
  • the modulating compounds which function as activators and inhibitors of PAR 2 proteins are small molecules.
  • the present invention rovides small molecule compounds encompassed within Formula I:
  • Formula I is not limited to a particular chemical moiety for R 1; R 2 , R3, and R4.
  • the particular chemical moiety for R 1; R 2 , R3, and R4 independently include any chemical moiety that permits the resulting compound to function as an inhibitor of PAR 2 protein activity.
  • the particular chemical moiety for R 1; R 2 , R3, and R4 independently include any chemical moiety that permits the resulting compound to function as an activator of PAR2 protein activity.
  • Ri is selected from
  • R2 is an amino acid selected from a Leu, He, Val, Cha, Arg, Orn, , Dap, Thr, Ser, and Tyr.
  • R3 is selected from 2-furoyl O ), 3- methylbutyryl
  • the modulating compound is the PAR 2 antagonist C391 (
  • compositions of the present invention are useful in treating conditions characterized with aberrant PAR 2 acitivty.
  • compositions comprising PAR 2 modulating compounds e.g., mimetic peptides, small molecules
  • Such conditions include, but are not limited to, asthma, chronic pain, cancer and/or vascular disorders.
  • the compositions and methods of the present invention are used to treat cells, tissues, organs, or pathological conditions and/or disease states in an animal (e.g.
  • a mammalian patient including, but not limited to, humans and veterinary animals
  • various diseases and pathologies are amenable to treatment or prophylaxis using the present methods and compositions.
  • a non-limiting exemplary list of these diseases and conditions includes, but is not limited to, cancers having aberrant activity, inflammatory conditions having aberrant PAR 2 activity, asthma, chronic pain, and/or vascular disorders having aberrant PAR 2 activity.
  • Some embodiments of the present invention provide methods for administering an effective amount of a PAR 2 modulating compound (e.g., mimetic peptide, small molecule) of the invention and at least one additional therapeutic agent (including, but not limited to, pain relieving agents, chemotherapeutic antineoplastics, apoptosis-modulating agents, antimicrobials, antivirals, antifungals, and anti-inflammatory agents) and/or therapeutic technique (e.g., surgical intervention, and/or radiotherapies).
  • a PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • additional therapeutic agent including, but not limited to, pain relieving agents, chemotherapeutic antineoplastics, apoptosis-modulating agents, antimicrobials, antivirals, antifungals, and anti-inflammatory agents
  • therapeutic technique e.g., surgical intervention, and/or radiotherapies.
  • anti-inflammatory agents include steroidal anti-inflammatory agents (e.g., albuterol), and non-steroidal anti-inflammatory agents.
  • anticancer agents are contemplated for use in the methods of the present invention. Indeed, the present invention contemplates, but is not limited to, administration of numerous anticancer agents such as: agents that induce apoptosis; polynucleotides (e.g. , anti- sense, ribozymes, siRNA); polypeptides (e.g. , enzymes and antibodies); biological mimetics;
  • alkaloids alkaloids; alkylating agents; antitumor antibiotics; antimetabolites; hormones; platinum
  • monoclonal or polyclonal antibodies e.g. , antibodies conjugated with anticancer drugs, toxins, defensins), toxins; radionuclides; biological response modifiers (e.g. , interferons (e.g. , IFN-a) and interleukins (e.g., IL-2)); adoptive immunotherapy agents; hematopoietic growth factors; agents that induce tumor cell differentiation (e.g. , all-trans-retinoic acid); gene therapy reagents (e.g.
  • antisense therapy reagents and nucleotides include tumor vaccines; angiogenesis inhibitors; proteosome inhibitors: NF-KB modulators; anti-CDK compounds; HDAC inhibitors; and the like.
  • angiogenesis inhibitors include angiogenesis inhibitors, anti-CDK compounds, HDAC inhibitors; and the like.
  • proteosome inhibitors include NF-KB modulators; anti-CDK compounds; HDAC inhibitors; and the like.
  • Numerous other examples of chemotherapeutic compounds and anticancer therapies suitable for coadministration with the disclosed compounds are known to those skilled in the art.
  • the pain relieving agents include, but are not limited to, analgesic drugs and respective antagonists.
  • analgesic drugs include, but are not limited to, paracetamol and Non-steroidal anti-inflammatory drugs (NSAIDs), COX-2 inhibitors, opiates and morphonimimetics, and specific analgesic agents.
  • NSAIDs include, but are not limited to, salicylates (e.g., Acetylsalicylic acid (Aspirin), Amoxiprin, Benorylate/Benorilate, Choline magnesium salicylate, Diflunisal,
  • salicylates e.g., Acetylsalicylic acid (Aspirin), Amoxiprin, Benorylate/Benorilate, Choline magnesium salicylate, Diflunisal,
  • Ethenzamide Faislamine, Methyl salicylate, Magnesium salicylate, Salicyl salicylate,
  • Salicylamide arylalkanoic acids
  • arylalkanoic acids e.g., Diclofenac, Aceclofenac, Acemethacin, Alclofenac, Bromfenac, Etodolac, Indometacin, Nabumetone, Oxametacin, Proglumetacin, Sulindac, Tolmetin
  • 2-arylpropionic acids e.g., Ibuprofen, Alminoprofen, Benoxaprofen, Carprofen,
  • COX-2 inhibitors include, but are not limited to Celecoxib, Etoricoxib, Lumiracoxib, Parecoxib, Rofecoxib, Valdecoxib.
  • opiates include, but are not limited to, natural opiates (e.g., alkaloids contained in the resin of the opium poppy including morphine, codeine and thebaine), semi-synthetic opiates (e.g., created from the natural opioids, such as hydromorphone, hydrocodone, oxycodone, oxymorphone, desomorphine, diacetylmorphine (Heroin), nicomorphine, dipropanoylmorphine, diamorphine, benzylmorphine, Buprenorphine, Nalbuphine, Pentazocine, meperidine, diamorphine, and ethylmorphine), fully synthetic opioids (e.g., such as fentanyl, pethidine, Oxycodone,
  • natural opioids such as hydromorphone, hydrocodone, oxycodone, oxymorphone, desomorphine, diacetylmorphine (Heroin), nicomorphine, dipropanoy
  • endorphins e.g., produced naturally in the body, such as endorphins, enkephalins, dynorphins, and endomorphins.
  • analgesics include, but are not limited to, tricyclic antidepressants (e.g., amitriptyline, carbamazepine, gabapentin, and pregabalin), Tetrahydrocannabinol, ketamine, clonidine, a 2 -adrenoreceptor agonists, mexiletine, Orphenadrine, cyclobenzaprine, scopolamine, atropine, gabapentin, first-generation antidepressants and other drugs possessing anticholinergic and/or antispasmodic.
  • tricyclic antidepressants e.g., amitriptyline, carbamazepine, gabapentin, and pregabalin
  • Tetrahydrocannabinol ketamine
  • clonidine a 2 -adrenoreceptor agonists
  • mexiletine e.g., mexiletine, Orphenadrine, cyclobenzaprine, sco
  • pain-relieving agents include anesthetic drugs.
  • anesthetic drugs include, but are not limited to, local anesthetics (e.g., procaine, amethocaine, cocaine, lidocaine, prilocaine, bupivacaine, levobupivacaine, ropivacaine, dibucaine), inhaled anesthetics (e.g., Desflurane, Enflurane, Halothane, Isoflurane, Nitrous oxide, Sevoflurane, Xenon), intravenous anesthetics (e.g., Barbiturates (e.g., amobarbital (Amytal), pentobarbital (Nembutal), secobarbital (Seconal), Phenobarbital, Methohexital, Thiopental, Methylphenobarbital, Metharbital, Barbexaclone)), Benzodiazepines (e.g., alpha, a
  • pain-relieving agents include anticonvulsant drugs.
  • anticonvulsant drugs include, but are not limited to, aldehydes (e.g., paraldehyde), aromatic allylic alcohols (e.g., stiripentol), barbiturates (e.g., amobarbital (Amytal), pentobarbital (Nembutal), secobarbital (Seconal), Phenobarbital, Methohexital, Thiopental, Methylphenobarbital, Metharbital, Barbexaclone), benzodiazepines (e.g., alprazolam, bromazepam (Lexotan), chlordiazepoxide (Librium), Clobazam, Clonazepam, Clorazepate, Diazepam, Midazolam, Lorazepam, Nitrazepam, temazepam, nimetazepam, Estazolam, Fl
  • pain-relieving agents include muscle relaxant drugs.
  • muscle relaxant drugs include, but are not limited to, depolarizing muscle relaxants (e.g., depolarizing muscle relaxants (e.g., depolarizing muscle relaxants), depolarizing muscle relaxants (e.g., depolarizing muscle relaxants, depolarizing muscle relaxants, depolarizing muscle relaxants, depolarizing muscle relaxants, depolarizing muscle relaxants, depolarizing muscle relaxants, depolarizing muscle relaxants (e.g., depolarizing muscle relaxants, e.g.
  • Succinylcholine short acting non-depolarizing muscle relaxants (e.g., Mivacurium,
  • Rapacuronium intermediate acting non-depolarizing muscle relaxants (e.g., Atracurium, Cisatracurium, Rocuronium, Vecuronium), and long acting non-depolarizing muscle relaxants (e.g., Alcuronium, Doxacurium, Gallamine, Metocurine, Pancuronium, Pipecuronium, d-Tubocurarine).
  • intermediate acting non-depolarizing muscle relaxants e.g., Atracurium, Cisatracurium, Rocuronium, Vecuronium
  • long acting non-depolarizing muscle relaxants e.g., Alcuronium, Doxacurium, Gallamine, Metocurine, Pancuronium, Pipecuronium, d-Tubocurarine.
  • a PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • one or more additional agents e.g., anti -inflammatory agents, anticancer agents, pain-relieving agents
  • an animal e.g., a human patient
  • additional agents e.g., anti -inflammatory agents, anticancer agents, pain-relieving agents
  • the PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • the one or more additional agents e.g., anti-inflammatory agents, anti-cancer agents, pain-relieving agents
  • additional agents e.g., anti-inflammatory agents, anti-cancer agents, pain-relieving agents
  • additional agents e.g., anti-inflammatory agents, anti-cancer agents, pain-relieving agents
  • the PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • the one or more additional agents e.g., anti -inflammatory agents, anti-cancer agents, pain-relieving agents
  • additional agents e.g., anti -inflammatory agents, anti-cancer agents, pain-relieving agents
  • the PAR 2 modulating compound (e.g., mimetic peptide, small molecule) and the additional agent are administered concurrently but on different schedules, e.g., the PAR 2 modulating compound (e.g., mimetic peptide, small molecule) is administered daily while the additional agent is administered once a week, once every two weeks, once every three weeks, or once every four weeks. In other embodiments, the PAR 2 modulating compound (e.g., mimetic peptide, small molecule) is administered once a week while the additional agent is administered daily, once a week, once every two weeks, once every three weeks, or once every four weeks.
  • the PAR 2 modulating compound (e.g., mimetic peptide, small molecule) is administered once a week while the additional agent is administered daily, once a week, once every two weeks, once every three weeks, or once every four weeks.
  • compositions within the scope of this invention include all compositions wherein the PAR 2 modulating compounds (e.g., mimetic peptides, small molecules) of the present invention are contained in an amount that is effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art.
  • the PAR 2 modulating compounds e.g., mimetic peptides, small molecules
  • about 0.01 to about 25 mg/kg is orally administered to treat, ameliorate, or prevent such disorders.
  • the dose is generally about one-half of the oral dose.
  • a suitable intramuscular dose would be about 0.0025 to about 25 mg/kg, or from about 0.01 to about 5 mg/kg.
  • the unit oral dose may comprise from about 0.01 to about 1000 mg, for example, about 0.1 to about 100 mg of the PAR2 modulating compound (e.g., mimetic peptide, small molecule).
  • the unit dose may be administered one or more times daily as one or more tablets or capsules each containing from about 0.1 to about 10 mg, conveniently about 0.25 to 50 mg of the PAR2
  • modulating compound e.g., mimetic peptide, small molecule
  • the PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • the PAR 2 modulating compound may be present at a concentration of about 0.01 to 100 mg per gram of carrier.
  • the PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • the PAR 2 modulating compound is present at a concentration of about 0.07-1.0 mg/ml, for example, about 0.1-0.5 mg/ml, and in one
  • the PAR 2 modulating compound e.g., mimetic peptide, small molecule
  • the PAR 2 modulating compounds may be administered as part of a pharmaceutical preparation containing suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the PAR 2 modulatign compounds into preparations which can be used
  • preparations particularly those preparations which can be administered orally or topically and which can be used for one type of administration, such as tablets, dragees, slow release lozenges and capsules, mouth rinses and mouth washes, gels, liquid suspensions, hair rinses, hair gels, shampoos and also preparations which can be administered rectally, such as suppositories, as well as suitable solutions for administration by intravenous infusion, injection, topically or orally, contain from about 0.01 to 99 percent, in one embodiment from about 0.25 to 75 percent of active mimetic peptide(s), together with the excipient.
  • compositions of the invention may be administered to any patient that may experience the beneficial effects of the PAR 2 modulating compounds (e.g., mimetic peptides, small molecules) of the invention.
  • PAR 2 modulating compounds e.g., mimetic peptides, small molecules
  • mammals e.g., humans, although the invention is not intended to be so limited.
  • Other patients include veterinary animals (cows, sheep, pigs, horses, dogs, cats and the like).
  • the PAR 2 modulating compounds e.g., mimetic peptides, small molecules
  • pharmaceutical compositions thereof may be administered by any means that achieve their intended purpose.
  • administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes.
  • administration may be by the oral route.
  • the dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
  • compositions of the present invention are manufactured in a manner that is itself known, for example, by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes.
  • pharmaceutical preparations for oral use can be obtained by combining the active mimetic peptides with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
  • Suitable excipients are, in particular, fillers such as saccharides, for example lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binders such as starch paste, using, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone.
  • fillers such as saccharides, for example lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binders such as starch paste, using, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose,
  • disintegrating agents may be added such as the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate.
  • Auxiliaries are, above all, flow-regulating agents and lubricants, for example, silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and/or polyethylene glycol.
  • Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices.
  • concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures.
  • suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate, are used.
  • Dye-stuffs or pigments may be added to the tablets or dragee coatings, for example, for identification or in order to characterize combinations of active mimetic peptide doses.
  • Other pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol.
  • the push-fit capsules can contain the active mimetic peptides in the form of granules that may be mixed with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active mimetic peptides are in one embodiment dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin.
  • stabilizers may be added.
  • Possible pharmaceutical preparations that can be used rectally include, for example, suppositories, which consist of a combination of one or more of the active mimetic peptides with a suppository base.
  • Suitable suppository bases are, for example, natural or synthetic triglycerides, or paraffin hydrocarbons.
  • gelatin rectal capsules that consist of a combination of the active mimetic peptides with a base.
  • Possible base materials include, for example, liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.
  • Suitable formulations for parenteral administration include aqueous solutions of the active mimetic peptides in water-soluble form, for example, water-soluble salts and alkaline solutions.
  • suspensions of the active mimetic peptides as appropriate oily injection suspensions may be administered.
  • Suitable lipophilic solvents or vehicles include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides or polyethylene glycol-400.
  • Aqueous injection suspensions may contain substances which increase the viscosity of the suspension include, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran.
  • the suspension may also contain stabilizers.
  • the topical compositions of this invention are formulated in one embodiment as oils, creams, lotions, ointments and the like by choice of appropriate carriers.
  • Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than C 12 ).
  • the carriers may be those in which the active ingredient is soluble.
  • Emulsifiers, stabilizers, humectants and antioxidants may also be included as well as agents imparting color or fragrance, if desired.
  • transdermal penetration enhancers can be employed in these topical formulations. Examples of such enhancers can be found in U.S. Pat. Nos. 3,989,816 and 4,444,762.
  • Ointments may be formulated by mixing a solution of the active ingredient in a vegetable oil such as almond oil with warm soft paraffin and allowing the mixture to cool.
  • a vegetable oil such as almond oil
  • a typical example of such an ointment is one that includes about 30% almond oil and about 70% white soft paraffin by weight.
  • Lotions may be conveniently prepared by dissolving the active ingredient, in a suitable high molecular weight alcohol such as propylene glycol or polyethylene glycol.
  • Protease-activated receptor-2 belongs to a four-member family of G-Protein coupled receptors (GPCRs) that contain internal ligands exposed following exogenous or endogenous protease cleavage of the extracellular amino terminus.
  • GPCRs G-Protein coupled receptors
  • PAR 2 is associated with a variety of inflammatory conditions, including asthma and pain.
  • the contributions of PAR 2 signalling to disease has been hindered by the lack of potent, efficacious antagonists, and their potential for biased-ligand signalling. It was recently demonstrated that lipid tethering of known PAR 2 peptidomimetic agonists based on the primary trypsin cleavage sequence (SLIGRL) increased their potency > 200 fold.
  • SLIGRL primary trypsin cleavage sequence
  • lipid tethering hexadecyl (Hdc) group with polyethylene glycol (PEG) spacers
  • Compound 562 (C562), 2-aininothiazol-4yl-SKGRS-P£G 5 -Hcfc blocks PAR 2 Ca 2+ signalling elicited via peptidomimetics (2-at-LIGRL-NH 2 ) or via asthma associated protease activation (Alternaria alternata filtrates) in cultured human bronchial epithelial cells (16HBE14o-).
  • This compound was a biased-signalling antagonist in that it had no effect on mitogen activated protein kinase (MAPK) signalling, the other major signalling pathway activated via PAR 2 .
  • MAPK mitogen activated protein kinase
  • C595 is closely related to the previously described potent and specific PAR 2 agonist, 2-at-LIGR-Pi?G3-Hcfc.
  • experiments screened a series of potential PAR 2 ligands with a heterocycle serine substitute followed by four amino acids (XXGR) and the PEG ⁇ -Hdc lipid tether.
  • C608 2-at-TIGR-Pi?G 3 -Hcfc
  • C608 blocked PAR 2 -dependent Ca 2+ signalling via protease or peptidomimetics without effects on MAPK signalling.
  • C562, C595 and C608 are novel pharmacological tools that can be used to evaluate the physiological consequences of PAR 2 full and biased ligand signalling.
  • Fig. 2 shows a schematic for PAR 2 signalling, and the measuring of PAR 2 signalling.
  • Fig. 3 shows PAR 2 tethered ligand probe development - trypsin site.
  • Fig. 4 shows PAR 2 tethered ligand probe development - kallikrein site.
  • Fig. 5 shows in vitro physiological PAR 2 agonist screening using xCELLigence.
  • Fig. 6 shows 2-at-TIGRJ ) £G 3 -H£fc signalling assays.
  • Tables 1, 2, 3, 4 and 5 provide additional PAR 2 mimetic peptide is configured to modulate PAR 2 biological activity.
  • kallikrein site directed tethered ligands are biased antagonists: a) RTCA signalling antagonists; b) Ca + signalling anatagonist; c) no effect on MAPK pathways.
  • C608 is a partial agonist and biased antagonist a) Partial RTCA agonist and potent anatagonist (low concentrations); b) Partial Ca 2+ signalling agonist and potent antagonist (low concentrations); c) Full MAPK agonist.

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Abstract

La présente invention concerne le domaine de la pharmacologie médicale. L'invention concerne notamment des composés modulant le récepteur activé par la protéase de type 2 (PAR2) (par exemple, des peptides mimétiques), des compositions comprenant ces composés de modulation et leur utilisation en tant qu'agents thérapeutiques pour traiter les états impliquant l'activité de PAR2.
PCT/US2017/025511 2016-04-01 2017-03-31 Peptides mimétiques par2 et utilisations desdits peptides WO2017173347A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019199800A1 (fr) * 2018-04-09 2019-10-17 Arizona Board Of Regents On Behalf Of The University Of Arizona Modulateurs de par2 à petites molécules et utilisations associées
WO2022117882A2 (fr) 2020-12-03 2022-06-09 Domain Therapeutics Nouveaux inhibiteurs de par-2
WO2023233033A1 (fr) 2022-06-03 2023-12-07 Domain Therapeutics Nouveaux inhibiteurs de par-2
US11925655B2 (en) 2014-06-13 2024-03-12 Bach Biosciences, Llc FAP-activated therapeutic agents, and uses related thereto
US12023316B2 (en) 2014-06-13 2024-07-02 Bach Biosciences, Llc FAP-activated therapeutic agents, and uses related thereto

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US20060104944A1 (en) * 2004-11-18 2006-05-18 Mousa Shaker A Activators and inhibitors of protease activated receptor2 (PAR2) and methods of use

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AU2002327323A1 (en) 2001-07-24 2003-02-17 High Throughput Genomics, Inc. Method and device for directed sort combinational synthesis

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US20060104944A1 (en) * 2004-11-18 2006-05-18 Mousa Shaker A Activators and inhibitors of protease activated receptor2 (PAR2) and methods of use

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FLYNN ET AL.: "Development of highly potent protease-activated receptor 2 agonists via synthetic lipid tethering", FASEB J., vol. 27, 2013, pages 1498 - 1510, XP055428718 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11925655B2 (en) 2014-06-13 2024-03-12 Bach Biosciences, Llc FAP-activated therapeutic agents, and uses related thereto
US12023316B2 (en) 2014-06-13 2024-07-02 Bach Biosciences, Llc FAP-activated therapeutic agents, and uses related thereto
WO2019199800A1 (fr) * 2018-04-09 2019-10-17 Arizona Board Of Regents On Behalf Of The University Of Arizona Modulateurs de par2 à petites molécules et utilisations associées
US11952379B2 (en) 2018-04-09 2024-04-09 Arizona Board Of Regents On Behalf Of The University Of Arizona Small molecule modulators of PAR2 and uses thereof
WO2022117882A2 (fr) 2020-12-03 2022-06-09 Domain Therapeutics Nouveaux inhibiteurs de par-2
WO2023233033A1 (fr) 2022-06-03 2023-12-07 Domain Therapeutics Nouveaux inhibiteurs de par-2

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